Difference between revisions of "User:Wesley J. Houston/Notebook/MAP/2013/03/18"

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(3/18/2013)
(3/18/2013)
Line 21: Line 21:
 
| &nbsp;            || 1    || 2    ||
 
| &nbsp;            || 1    || 2    ||
 
|-
 
|-
| Insert DNA (KAH182)|| 6.0  || ---  ||
+
| Insert DNA (KAH182)|| 8.0  || ---  ||
 
|-
 
|-
 
| Vector DNA (KAH201)|| 1.0  || 1.0  ||
 
| Vector DNA (KAH201)|| 1.0  || 1.0  ||
 
|-
 
|-
| 2x lgn buf (Roche) || 8.0  || 5.0  ||
+
| 2x lgn buf (Roche) || 10.0  || 5.0  ||
 
|-
 
|-
 
| T4 ligase (NEB)    || 1.0  || 1.0  ||
 
| T4 ligase (NEB)    || 1.0  || 1.0  ||
Line 31: Line 31:
 
| dH<sub>2</sub>O    ||  0  || 3.0  ||
 
| dH<sub>2</sub>O    ||  0  || 3.0  ||
 
|-
 
|-
| &nbsp;            || 16 μL || 10 μL ||
+
| &nbsp;            || 20 μL || 10 μL ||
 
|}
 
|}
  

Revision as of 12:32, 18 March 2013

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3/18/2013

  • Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675

> Ligations, following corrected version from 3/4/13, with 50 ng of vector instead of 25 ng.

Ligation Plate results (lig : neg crtl) 03/18/13
1. KAH201 (S/P)/3403, 50 ng + KAH182 (X/P)/1675, 50 ng TBD
2. KAH201 (S/P)/3403, 50 ng TBD
  1 2
Insert DNA (KAH182) 8.0 ---
Vector DNA (KAH201) 1.0 1.0
2x lgn buf (Roche) 10.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 0 3.0
  20 μL 10 μL
  • Follow quick transformation protocol
  • Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar